S-(&#39; -substitutedamino-ethyl)-cysteine

ABSTRACT

New basic derivatives of cysteine, having a favourable action on epithelia as well as on their secretions, are characterised by carrying a (tertiary amino)-ethyl group or a (tertiary amino)benzyl group attached to the sulphur atom of cysteine.

United States Patent [191 J oullie et al.

[451 Jul 22, 1975 S-( B-SUBSTlTUTEDAMINO-ETl-IYL CYSTEINE [75] Inventors: Maurice Joullie,

St-Germain-en-Laye, France; Lucien Lakah, Paris; Gabriel F. Maillard, Paris; Pierre Muller, Paris, all of France [73] Assignee: Recherches Pharmaceutiques et Scientifiques, Paris, France [22] Filed: Oct. 2, 1972 [21] Appl. No.2 294,105

[30] Foreign Application Priority Data Oct. 4, 1971 France 71.35680 [58] Field of Search... 260/247.1 R, 293.85, 326.84, 260/268 R, 534 S, 326.2

[56] References Cited OTHER PUBLICATIONS Hope et al., J. Chem. Soc. (C), 1098-1101.

Hermann et al., C.A., 72: 67238p, (1970).'

Primary Examiner-Lorraine A. Weinberger Assistant ExaminerMichael Shippen Attorney, Agent, or Firm-Stevens, Davis, Miller & Mosher [5 7] ABSTRACT New basic derivatives of cysteine, having a favourable action on epithelia as well as on their secretions, are characterised by carrying a (tertiary amino)-ethyl group or a (tertiary amino)-benzyl group attached to the sulphur atom of cysteine.

2 Claims, No Drawings S-( B-SUBSTITUTEDAMINO-ETHYL )-CYSTEINE The invention relates to new basic derivatives of. cysteine and pharmacologically acceptable acid addition salts thereof, as well as to their preparation and their applications.

Its purpose is especially to provide new sulphurcontaining compounds which are capable of having a favourable action on epithelia as well as on their secretions.

The new compounds are characterised in that they carry a tertiary amine radical at the end of a carbon chain bonded to the sulphur atom of cysteine, specifically a (tertiary amin)-ethyl group, the tertiary amino group being the radical of dimethylamine, diethylamine, pyrrolidine, piperidine, morpholine or 4-methy1- piperazine, or a (tertiary amino)-benzyl group, the tertiary amino group being the radical of pyrrolidine, piperidine, morpholine or 4-methyl-piperazine.

In order to prepare these (tertiary-amine)-type compounds, it is possible, according to the'invention, to react cysteine in the form of its hydrochloride with a (tertiary amino)-ethyl or (tertiary amino)-benzyl halide in the form of the hydrohalide, the halogen being chlorine or bromine, in the presence of an alkali metal hydroxide, in an aqueous medium.

The following Examples illustrate the invention.

EXAMPLE 1 S-(B-dimethylamino-ethyl)-cysteine (LJ 222) and its monohydrochloride (LJ 388) 70 g (0.4 mol) of cysteine hydrochloride monohydrate in 100 ml of freshly distilled water are placed in a flask equipped with a stirrer and a nitrogen inlet.

48 g (1.2 mol) of sodium hydroxide dissolved in 100 ml of freshly distilled water are added gradually to the solution obtained.

The temperature is increased to about 60 and 120 g (approximately 30% excess) of fi-dimethylaminobromoethane hydrobromide are introduced over the course of 20 minutes.

The solution obtained is passed over a Dowex 50 resin and eluted with ammonia solution containing 33% by volume of NH Ol-l.

The elution product is distilled in vacuo to dryness, the residue is dried in vacuo in the presence of a dehydrating agent and the product is finally recrystallised from absolute ethanol. 49 g (64% yield) of it are obtained.

Melting point 212 213; isoelectric point 6.32 1

' Analysis gives for C-,H O N S (molecular weight calculated C g 43.75 found 43.80 4 H 8.33 8.70 N 14.58 14.10 S 16.66

chloric acid is added, the solution is brought to dryness under 15 mm of mercury and the residue is dried azeotropically as a suspension in benzene and then the product is dried in vacuo.

EXAMPLE 2 S-(B-diethylamino-ethyl)-cysteine (LJ 427):

This product is prepared in accordance with the technique of Example 1 from:

43.52 g of B-diethylamino-chloroethane hydrochloride (0.23 mol 10% excess) and 40.36 g of cysteine hydrochloride monohydrate (0.23 mol).

15.9 g (31% yield) of it are obtained.

Melting point approximately 215;

Isoelectric point 8.99

EXAMPLE 3 S-(B-pyrrolidino-ethyl)-cysteine (LJ 428) and calculated C 51.72 found 51.73 H 8.63 8.69 N 12.07 11.70

isoelectric point 8.97 Rf 0.534

EXAMPLE 4 S-(B-morpholino-ethyl)-cysteine (LJ 223) and its monohydrochloride (LJ 394) The same technique is applied with 35.1 g (0.2 mol) of cysteine hydrochloride monohydrate in 60 ml of distilled water,

24 g (0.6 mol) of NaOH in 60 m1 of distilled water and 39 g (0.2 mol 5% excess) of B-chloroethylmorpholine.

After passing over a Dowex 50 resin, the product is recrystallised from a mixture of 200 ml of ethanol and 30 ml of water.

22 g (yield: 47%) of it are obtained.

Melting point 237 238 (instantaneous block) or 205 206 (capillary tube), isoelectric point 6.56

Analysis gives for C H O N S (molecular weight calculated C 46.15 found 45.75

EXAMPLE S-(B-piperidino-ethyl)-cysteine (LJ 225) and its monohydrochloride (LJ 393).

CH -CH H C CHz-CHKNCH CH SCH CHCOOH The same technique is applied with:

57 g of cysteine hydrochloride monohydrate (0.325 mol),

40 g of NaOH (1 mol) and 66 g of B-piperidino-chloroethane hydrochloride (0.358 mol).

The crude product is passed over 1 kg of Dowex resin and recrystallised twice from 96% strength ethanol.

35 g (yield: 40%) of it are obtained.

Melting point 216-2l7; isoelectric point 8.88

Analysis gives for C H O N S (molecular weight 232) calculated C 5l.72 found 51.73

EXAMPLE 6 S-[/3-(4-methyl-piperazino)-ethyl]-cysteine (L.J. 541):

The same technique is applied with 22.35 g of cysteine hydrochloride monohydrate (0.273 mol) in 100 ml of distilled water, 21.4 g of NaOH (0.534 mol) and 33 g of l-methyl- 4-(B-chloroethyl)-piperazine dihydrochloride (0.140 mol).

The mixture is heated for l hour at 60, 9 g of the same hydrochloride excess) are then added and the mixture is heated for 1 hour at 80 and then for 3 hours at between 60 and 70.

The solution is treated with Dowex 50 resin and recrystallised from ethanol.

Melting point 205-206; isoelectric point 8.30

The compounds of Examples 1 to 6 were subjected to pharmacological experiments, the results of which are given below.

Toxicology The maximum doses tolerated (MDT) in mice, during oral administration, are given in Table 1 below, the compounds having first been neutralised, because of their high basicity, by means of hydrochloric acid; the compound LJ 541 was administered intraperitoneally and it is its LD 50 which is given.

TABLE 1 ACUTE TOXICITY u 388 MDT 25 g/kg LJ 225 MDT 4 g/kg LJ 394 MDT as g/kg [u 394 LD 50, i .v.8.2 g/kg .u 427 MDT 55 g/kg u 428 MDT ;5 g/kg u 541 MDT 25 g/kg Since the products are very basic, they are neutralised before the toxicity experiment by means of 2 N HCl.

Effect on the bronchial epithelium and its secretion In order to create experimentally a bronchial hypersecretion, male rats of 200 g, in a leakproof cage, are daily made to inhale S0 at a concentration of 300 ppm, for 2 to 5 hours per day and are made to do so for 5 days per week.

After approximately 120 hours of inhalation, an endobronchial retention of mucus is observed.

After killing the animals by cutting the abdominal aorta, under nembutal-induced anaesthesia, the left lung is removed, fixed in formaldehyde and out along the main bronchus after endotracheal colouration by means of Alcian Blue. The bronchus is examined under binocular magnification, in an aqueous medium, and for each lung, the presence or absence of bronchial retention is noted, as well as its appearance (compact plug, nodular agmen, blistery agglomeration and the like).

A quantitative and qualitative estimate of a secretion of mucus in the various batches is thus obtained.

A left lung is fixed in Dubosq-Brasil liquid, set in paraffin and cut in such a way that the complete bronchial tree, namely the main bronchus with four branches, can be spread on a histological slide. The muciparous cells are coloured by means of PAS (periodic acid Schiff) in accordance with the l-lotchkiss-Mac Magnus technique.

Thiol groups are detected by 6,6-dihydroxydinaphthyl 2,2'-disulphide in the bronchial mucus, on the histological sections thus prepared.

100 male rats of 200 g were subjected to repeated inhalations of $0 in accordance with the technique described above. From the start of the experiment, three animals which showed hypersensitivity to were removed. After 80 hours exposure to the irritant gas,

seven rats were killed and autopsies were carried out on them to verify bronchial hypersecretion.

From this time, the 90 remaining animals were di- Gelatine-coated pills:

LJ 394 vrded into nine batches of and, whilst being sub- Talc 86322:; 832% jected to the inhalation sessions, they were given re- 5 gf t d f 8 0-002 spectively: g e-coa e p 0 g or 0.250

gatcl}: gljtllggdswfltll 1 ml The gelatine-coated pills are prepared by dividing the 2:2 I 427 500 mg/kg' 10 mixture of the above powders by means of a machine Batch 4; LJ. 428 administered orall of the usual type for fillmg gelatme-coated pills.

Batch 5: LJ. 223 (HCl) in a volume of 1 ml Batch 6: LJ. 225

Batch 7: LJ. 541

daily, 5 days per week, for 16 days, which corresponds Tablets,

to 110 hours of inhalation of S0 U 225 o 400 The number of ammals showing endobronchlal re- Silica gel 0:010 g tention of mucus is shown in the following table: Stearic acid 0.020 g Wheat starch 0.045 g TABLE II 20 for a completed tablet containing 475 mg.

Controls, SO; alone: 8/10 The tablets are manufactured by direct compression 4/10 of the mixture of the constituents in a machine of the LJ. 427: 4/10 usual type. Dragee-type tablets:

5/10 The above tablets can be made into dragees by the conventional methods by coating with sugar, using a A decrease in the caliciform hyperplasia, demonf i i l d h trated by colouration with P A S is observed with all e co'fmng conslsts 9 mixtures o pow ers Sue as d t d gum arabrc, starch, calcium carbonate, talc and sucrose enva Ives S u 16 The addition of these excipients brings the dragees to At the level of the bronchial mucus, a disappearance a final weight of 700 mg. of the SH groups is observed only in the batch of controls irritated by S0 In all the other batches, wherein the animals are treated with one of the new derivatives of cysteine, SH groups are present. suppositories;

It is obvious from what has been stated above that the L 3 3 0 200 0 0 new derivatives of cysteine have a favourable action on i i 28 g the secretion of bronchial epithelium, bringing the Suppository excipient, q.s.p. l g or 1.500 g amount of mucus produced back to the normal level 40 and restoring biochemical equilibrium to a perturbed bronchial Wall- The suppository excipient is either cacao butter or,

These new deflvahves Consequently he ee as preferably, a fatty acid glyceride such as described in active principles of medicines, particularly medicines the French Codex f 1965 under h h di S ifor oral administration in pneumology (bronchalsynthetic glycerides pages 527 to 529 P p y and acute and Chrome brehehlhs) and 1h The suppositories are manufactured in accordance oto-rhino-laryngology (acute sinusitis, acute otitrs and i h f ll i h d; rhino-pharyngitis). a. mixing of the S-(amino-alkyl)-cysteine with the In each of the examples given below, any one of the l d m yD- y derivatives other than that b. incorporation of this mixture into the melted exwhich is stated, can be used. cipient and casting in suitable moulds.

Drinkable solutions:

a. Drops:

Granules:

LJ 223 10 g Concentrated solution to be administered as drops: Colloidal silica gel 0.5 g Sucrose 88.5 g U 428 15 g Polyvinylpyrrolidone l g Glucose 10 g Methyl para-hydroxy-benzoate 0.15 g

Distilled water, q.s.p. 100 g For example, the LJ 223, the silica gel and the sucrose are mixed in a mixer-mill of the usual type. The mixture is moistened with a 5% strength aqueous solu- The methyl para-hydroxy-benzoate is dissolved at 80 tion of polyvinylpyrrolidone until a mass is obtained in water, and then the compound LJ 394 and the gluwhich is granulated and dried in an oven.

After drying, it is possible to perfume the granules by spraying an essence, for example, orange essence.

cose are added.The volume is made up to the desired level and the solution is filtered.

b. Syrup:

LJ 394 2 g or 5 g Methyl para-hydroxy-benzoate 0.15 g 0.l5 g Sucrose 50 to 7".) g 50 to 70 g Distilled water q.s.p. 100 g l00 g The methyl para-hydroxy-benzoate is dissolved in water at 80 and then the morpholino-ethyl-cysteine hydrochloride (LJ 394) is added and, when it is dissolved, the sucrose is added When the whole is limpid, it is filtered and divided into flasks.

The syrup can be scented with, for example, raspberry or cherry concentrate or any other scent.

The syrup can be coloured with a pharmaceutically tolerated dyestuff such as tartrazine.

The new derivatives can also be used in dermatology, (illnesses of exposed parts of the body, excess perspiration and seborrhea), in gynaecology (trophic disorders of the vaginal mucous membrane, vaginitis and cervicitis) and in cosmetology, by virtue of the trophic action of these derivatives, especially the restoration of the SH groups of the damaged mucous membrane, on epithelia such as those of the vagina and of the skin.

The following examples illustrate the invention in these applications.

Dennatological cream for the skin:

Glyceryl monostearate Diethylene glycol monostearate Paraffin oil Sweet almond oil lsopropyl myristate Poly-oxy-ethylene sorbitan oleate Pol -oxy-ethylene sorbitan stearate Sor ic acid Water scent q.s.p.

'Cream for the treatment of nails:

Cholesterol Lanolin Sorbitol Glyceryl stearate Poly-oxy-ethylene sorbitan oleate Sorbic acid Water, q.s.p.

Scent dyestuffs, q.s.

89 Ml)! Us Nae untmuennan on Working at 60, the glyceryl monostearate and the lanolin, previously melted at 65, are added to a solution, prepared under hot conditions, of sorbic acid in water to which the S-(morpholino-ethyl)-cysteine salt, the cholesterol, the sorbitol, the poly-oxy-ethylene sorbitan oleate and the dyestuff have been added.

The mixture is stirred until cold and then the scent is added.

Cleansing and treatment milk:

LJ 427 2 Lanolin oil 2 Propylene glycol 10 Propylene glycol monostearate S Vaseline oil 2 Sodium lauryl-sulphate 1. Methyl para-hydroxy-benzoate 0. Water, q.s.p. Scent, q.s.

Lotion for the scalp:

S-(B-pyrrolidino-ethyl)-cysteine, LJ 428 Caster oil Propylene glycol Ethyl alcohol Distilled water, q.s.p. Scent, q.s.

Preparation by simple solution. Billiantine liquid:

LJ 225 5 g Castor oil 3 g Diethylene glycol 5 g Ethyl alcohol 0 g Distilled water, q.s.p. 0 g

Preparation by simple solution. Brilliantine cream:

1...! 428 5 Castor oil 3 Perhydrosqualene 3 Poly-oxy-ethylene glycol stearate l Poly-oxy-ethylene glycol oleate l Sorbic acid 0 Water, q.s.p. 0 Scent, q.s.

in order to prepare the brilliantine, the sorbic acid is dissolved at in the water and the cysteine derivative is added, after which the poly-oxy-ethylene glycol stearate and oleate are added, followed by the perhydrosqualene and the castor oil. The mixture is left to cool and is perfumed. Tablets for gynaecology:

a) L]. 394 0.150 g for a table Lactose 1.350 g of 4 g Granulated excipient 2.500 g The granulated excipient is mixed with the other powders and the mixture is compressed.

b) L.Jv 428 0.100 g Boric acid 0.050 g for a tablet of Granulated excipient 2 350 g 2.5 g Lavender essence 0 1 ml The granulated excipient has the following composition:

The LJ. 393 is mixed with the lactose and incorporated into the excipient at 37.

b) L]. 427 .100 g Monosodium phosphate 0.200 g for an pill of Sodium bicarbonate 0.200 g 3 g lmhausen excipient 2.500 g In the different examples proposed, it is possible to envisage combinations with local antiseptics, bactericidal agents, anti-fungal agents, and in the case of vaginal pills, trichomonocides.

Deodorant lotion (underarms) Boric acid 0.200 g Propylene glycol 1.000 g Camphor (10% strength solution in alcohol) 0.l00 g Hexachlorophene 0.500 g 95% strength ethyl alcohol 62.000 g L]. 223 2.000 g Perfume and preservative, q.s.

We claim:

1. A cysteine derivative selected from the group consisting of S-(B-dimethylamino-ethyl)-cysteine, S-(B- diethylaminoethyl)-cysteine, S-(B-pyrrolidino-ethyhcysteine, S-( B-morpholinoethyl )-cysteine, S-( B- piperidino-ethyl )-cysteine and S- B-( 4-methylpiperazino)-ethyl]-cysteine or a monohydrochloride thereof.

2. The cysteine derivative S-(B-morpholino-ethyhcysteine or its monohydrochloride. 

1. A CYSTEINE DERIVATIVE SELECTED FROM THE GROUP CONSISTING OF S-(B-DIMETHYLAMINO-ETHYL)-CYSTEINE, S-(B-DIETHYLAMINOETHYL)-CYSTEINE, S-(B-PYRROLIDONO-ETHYL)-CYSTEINE, S-(B-MORPHOLINOETHYL)-CYSTEINE, S-(B-PIPERIDINO-ETHYL)-CYSTEINE AND S-(B-(4-METHYLPIPERAZINO)-ETHYL)-CYSTEINE OR A MONOHYDROCHLORIDE THEREOF.
 2. The cysteine derivative S-( Beta -morpholino-ethyl)-cysteine or its monohydrochloride. 